Vinexin ß Interacts with Hepatitis C Virus NS5A, Modulating Its Hyperphosphorylation To Regulate Viral Propagation.

UNLABELLED: Hepatitis C virus (HCV) nonstructural protein 5A (NS5A) is essential for HCV genome replication and virion production and is involved in the regulation of multiple host signaling pathways. As a proline-rich protein, NS5A is capable of interacting with various host proteins containing Src homology 3 (SH3) domains. Previous studies have suggested that vinexin, a member of the sorbin homology (SoHo) adaptor family, might be a potential binding partner of NS5A by yeast two-hybrid screening. However, firm evidence for this interaction is lacking, and the significance of vinexin in the HCV life cycle remains unclear. In this study, we demonstrated that endogenously and exogenously expressed vinexin ß coimmunoprecipitated with NS5A derived from different HCV genotypes. Two residues, tryptophan (W307) and tyrosine (Y325), in the third SH3 domain of vinexin ß and conserved Pro-X-X-Pro-X-Arg motifs at the C terminus of NS5A were indispensable for the vinexin-NS5A interaction. Furthermore, downregulation of endogenous vinexin ß significantly suppressed NS5A hyperphosphorylation and decreased HCV replication, which could be rescued by expressing a vinexin ß short hairpin RNA-resistant mutant. We also found that vinexin ß modulated the hyperphosphorylation of NS5A in a casein kinase 1α-dependent on manner. Taken together, our findings suggest that vinexin ß modulates NS5A phosphorylation via its interaction with NS5A, thereby regulating HCV replication, implicating vinexin ß in the viral life cycle. IMPORTANCE: Hepatitis C virus (HCV) nonstructural protein NS5A is a phosphoprotein, and its phosphorylation states are usually modulated by host kinases and other viral nonstructural elements. Additionally, cellular factors containing Src homology 3 (SH3) domains have been reported to interact with proline-rich regions of NS5A. However, it is unclear whether there are any relationships between NS5A phosphorylation and the NS5A-SH3 interaction, and little is known about the significance of this interaction in the HCV life cycle. In this work, we demonstrate that vinexin ß modulates NS5A hyperphosphorylation through the NS5A-vinexin ß interaction. Hyperphosphorylated NS5A induced by vinexin ß is casein kinase 1α dependent and is also crucial for HCV propagation. Overall, our findings not only elucidate the relationships between NS5A phosphorylation and the NS5A-SH3 interaction but also shed new mechanistic insight on Flaviviridae NS5A (NS5) phosphorylation. We believe that our results may afford the potential to offer an antiviral therapeutic strategy.

Microstructure and Texture Characterization of Duplex Stainless Steel Joints Welded by Alternating Magnetic Field-Assisted Hybrid Laser-GMAW Welding.

In this study, 2205 duplex stainless steel with 12 mm thickness was welded by alternating magnetic field-assisted laser arc hybrid welding. The effect of an alternating magnetic field on the proportion distribution of two phases of the welded joint was investigated. The texture distribution, grain boundary misorientation, and grain size of welded joints were analyzed and characterized. The uniform distribution of alloying elements in the two phases was improved by a 20 mT alternating magnetic field. The diffusion dissolution of Ni and N elements into the γ phase was promoted, which was conducive to the transition from the α to γ phase and reduced the precipitation of Cr2N, such that the ratio of γ to α was 43.4:56.6. The ratio of the two phases of the weld was balanced by the alternating magnetic field of 30 mT, such that the ratio of γ and α was 44.6:55.4 and the texture dispersion was weakened. The Σ3 twinning boundary of the austenite phase in the weld was transformed to HABs, the ferrite phase underwent dynamic recrystallization, and the austenite phase had a cube texture, copper texture, and goss texture.

Comparison of perioperative clinicopathologic outcome and postoperative survival of laparoscopic and open sphincter-sparing surgery in patients with rectal cancer: a retrospective study.

PURPOSE: To investigate the perioperative clinicopathologic outcome and postoperative survival of sphincter-sparing surgery by laparoscopic and open approach for patients with rectal cancer. Μethods: From January 1, 2008 to December 31, 2011, laparoscopic sphincter-sparing surgery and open sphincter-sparing surgery was performed in 228 patients with rectal cancer who were included in this study as open group (N=112) and laparoscopic group (N=116), respectively. The average follow-up time was approximately 5 years. RESULTS: Spearman's test showed that there was a slight negative correlation in overall survival and American Society of Anesthesiologists (ASA) grade (Spearman's r=-0.146, p=0.028), History of abdominal surgery (Spearman's r=-0.134, p=0.044) of all patients was statistically significant. There was no significant difference in survival between laparoscopic and open group (p=0.988). Kaplan-Meier curves showed that the total overall survival rates after laparoscopic and open sphincter-sparing surgery were similar in both groups. Log rank test showed that there were significant differences in overall survival among different ypTNM stages (pathological TNM after neoadjuvant chemotherapy) (p=0.002) and Charlson comorbidity index (p=0.03). CONCLUSIONS: Compared with open approach, laparoscopic sphincter-sparing surgery of rectal cancer had less intraoperative bleeding, less postoperative complications and faster recovery of intestinal function after operation. Survival of open surgery and laparoscopic rectal sphincter preservation surgery was similar in both groups. ypTNM stage and Charlson comorbidity index are the risk factors affecting the survival of patients with rectal cancer.